• Journal of Microbiology and Biotechnology
• /
• v.28 no.10
• /
• pp.1749-1759
• /
• 2018

Recombinant (rec) prion protein (PrP) is an extremely useful resource for studying protein misfolding and subsequent protein aggregation events. Here, we report mass production of high-purity rec-polypeptide encoding the C-terminal globular domain of PrP; (90-230) for human and (89-231) for murine PrP. These proteins were expressed as His-tagged fusion proteins in E. coli cultured by a high cell-density aerobic fermentation method. RecPrPs recovered from inclusion bodies were slowly refolded under reducing conditions. Purification was performed by a sequence of metal-affinity, cation-exchange, and reverse-phase chromatography. The current procedure yielded several dozens of milligrams of recPrP per liter with >95% purity. The purified recPrPs predominantly adopted an ${\alpha}$-helix-rich conformation and were functionally sufficient as substrates to measure the seeding activity of human and animal prions. Establishment of a procedure for high-level production of high-purity recPrP supports the advancement of in vitro investigations of PrP including diagnosis for prion diseases.

• Journal of Plant Biotechnology
• /
• v.29 no.4
• /
• pp.229-233
• /
• 2002

Ferritin is ubiquitous in bacteria, animals and plants. Ferritin is thought to play two main roles in living cells to provide iron for the synthesis of iron protein such as ferretoxin and cytochromes and to prevent damage from radicals produced by iron/dioxygen interaction. To enhance the resistance of potato to Erwinia carotovora, the soybean ferritin gene was introduced into the potato either under CaMV 35S or hsr203J promoter. Potato transgenic plants were screened by PCR analysis using specific primers to the ferritin gene. Expression of ferritin gene under CaMV 35S and hsr203J promoter in potato transgenic plants was confirmed by northern blot analysis. hsr203J promoter known to pathogen inducible in tobacco drives the induction upon Phytophthora infestan in potato and the transcript level of ferritin gene was extremely high after 24 hours post inoculation. One of transformants under CaMV 35S promoter was increased 2.5 fold than untransformant. Each one of transgenic potato containing gene promoter CaMV 35S and hsr203J-ferrtin fusion exhibited tolerance against potato soft rot.

• Fashion & Textile Research Journal
• /
• v.9 no.5
• /
• pp.538-544
• /
• 2007

We usually try to accomodate ourselves to our nature surroundings interacting with nature. There fore I've decided to apply nature materials, esp. the pine to Hair Art as a way of expressing our feelings from Nature. 'Hair Art' is a follow-up study based on the various artistic point of view. There has been various works of 'Hair Art' with nature materials. However, it is rare to see a work of 'Hair Art' with the pine. That's why I've decided to study more about this. This study is a new experiment in 'Hair Art' against a conventional idea "Hair Art values practicability." Therefore, the primary goal of this study is to do much for the cause of learning in 'Hair Art' and encourages some development of 'Hair Art' industry. I've researched an image of a pine tree in art of Joseon Dynasty and contemporary art. Then I've tried to apply the image to a work of 'Hair Art'. First of all, in a view of expression technique for the symbolism of the pine, the artists in Joseon Dynasty drew pictures of a pine tree with a paintbrush but the modern artists make a new attempt from the thought of Modernism. We can find it in some photos. Next, to express traditional oriental idea such as 'unconventional and elegance', comtemporary artists chose the symbolism of the pine tree as an object of their works like pen and ink sketches from the thought of Modernism. Third, in a fusion style picture which contains features of both oriental paintings and western paintings and in a sexualism style picture that depicts a harmony of a male and a female as a shape of a pine, we can find colorful images of a pine tree and Their figurative beauty in art. Those are another symbols of the pine. In conclusion, the implication of the pine tree still hasn't changed even there are differences of drawings of pine tree in the past and the present. I've tried to combine these symbolic ideas of the pine with 'Hair Art' and made 5 hair styles. Throughout the process of researching this topic which is 'The Study of Hair Art Using The Symbolism of The Pine', I've realized that pine trees make it possible to express intrinsic tough spirit of human being and abundance in color and figurative beauty in art. I hope this can contribute to the field of 'Hair Art' and would become an important educational resource for further study.

• Journal of Life Science
• /
• v.14 no.2
• /
• pp.291-295
• /
• 2004

Rooibos (Aspalathus linearis) (RB) is a leguminous shrub native to the mountainous areas of the northwestern Cape Province in South Africa. RB tea infusions are the fermentation products of its leaves and fine sterns, and known to have a high antioxidative activity due to the presence of flavonoids and phenolic acids. We investigated the effects of RB tea on the alleviation of oxidative stress on cultured rat cortical neurons in a hypoxic model. Measurement of lactate dehydrogenase (LDH) released into culture media revealed that RB increased cell viabilities in both normoxia (6-18%) and hypoxia (2-24%) dose-dependently (10-100 $\mu\textrm{g}$/ml) on 16 days in vitro (3 days after treatment). Visualization of cell morphology by expression of GFP-Hsc70 fusion protein showed that RB (50 $\mu\textrm{g}$/ml) reduced the average vacuolated soma from 55.4$\pm$4.59% (no RB addition) to 40.9$\pm$6.3% (RB addition) on 5 days after hypoxia. Our results proves efficacy of RB in the neuroprotection of hypoxic neurons and extend application for RB into the prevention and/or treatment of neuronal damages.

• Journal of Life Science
• /
• v.14 no.2
• /
• pp.325-330
• /
• 2004

The RAD3 gene of Saccharomyces cerevisiae is required for excision repair and is essential for cell viability. RAD3 encoded protein possesses a single stranded DNA-dependent ATPase and DNA and DNA-RNA helicase activities. To examine the extent of conservation of structure and function of RAD3 during eukaryotic evolution, the RAD3 homolog gene was isolated by screening of genomic DNA library. The isolated gene was designated as HRD3 (Homologue of RAD3 gene). The over-expressed HRD3 protein was estimated to be a 75 kDa in size which is in good agreement with the estimated by the nucleotide sequence of the cloned gene. Two-dimensional gel electrophoresis showed that a number of other protein spots dramatically disappeared when the HRD3 protein was overexpressed. The overexpressed RAD3 protein showed a toxicity in E. coli host, suggesting that this protein may be involved in the inhibition of protein synthesis and/or degradation of host protein. To determine which part of HRD3 gene contributes to the toxicity in E. coli, various fusion plasmids containing a partial sequence of HRD3 and lac'Z gene were constructed. These results suggest that the C-terminal domain of HRD3 protein may be important for both toxic effect in E. coli and for its role in DNA repair in S. pombe.

• Journal of Life Science
• /
• v.17 no.1 s.81
• /
• pp.56-63
• /
• 2007

Insulin stimulates the fusion of intracellular vesicles containing glucose transporter 4 (GLUT4) with plasma membrane in adipocytes and muscle cells. Here we show that adipocyte differentiation results in enhanced insulin sensitivity of glucose uptake. On the other hand, glucose uptake in response to platelet-derived growth factor (PDGF) stimulation was markedly reduced by adipocyte differentiation. Expression level of insulin receptor and caveolin-1 was dramatically increased during adipocyte differentiation. Adipocyte differentiation caused :ilightly enhanced activation of acutely transforming retrovirus AKT8 in rodent T cell lymphoma (Akt) by insulin stimulation. However, activation of Akt by PDGF stimulation was largely reduced. Activation of ERK was not detected in both fibroblasts and adipocytes after stimulation with insulin. PDGF-dependent activation of ERK was reduced by adipocyte differentiation. Insulin-dependent glucose uptake was abrogated by LY294002, a phosphatidylinositol 3-kinase (PI3K) inhibitor, in both fibroblasts and adipocytes. Also disassembly of caveolae structure by $methyl-\beta-cyclodextrin$ caused impairment of Akt activation and glucose uptake. Finally, insulin receptor, Akt, SH2-domain-containing inositol 5-phosphatase 2 (SHIP2), and regulatory subunit of PI3K are localized at lipid raft domain and the translocation was facilitated upon insulin stimulation. Given these results, we suggest that lipid raft provide proper site for insulin action for glucose uptake.

• Spatial Information Research
• /
• v.22 no.5
• /
• pp.27-41
• /
• 2014

Recently, BIM(Building Information Modeling) has been applied to the infrastructure such as road and bridge, and information about the outside environment of buildings is needed for maintaining and managing the large urban facilities. In addition, the convergence between spatial information and Big-data has a large potentiality, in respect that considerable profits and developments in other application problems such as various simulations and urban plans, national land security, may be brought about on the basis of the interoperability of information between BIM and GIS. Therefore, this study attempted to suggest the development direction of a model integrating building for spatial information analysis and city on the subject by comparing and analyzing difference between information system and shape expression of IFC, CityGML and LandXML to efficiently link information between IFC as a standard model of BIM and CityGML as a standard model in the GIS sector and to prepare a basic fusion strategy and a method of utilization between BIM and GIS. The result of the study are as follow. Firstly, contents and structure of IFC, CityGML and LandXML are compared and analyzed. Secondly, the development direction of iBUM(Integrated Building and Urban Model) suggested, which is based on convergence technology for analysis of space information. Finally, a strategy and method of the BIM and GIS are proposed in the iBUM environment.

• Journal of Life Science
• /
• v.16 no.7 s.80
• /
• pp.1199-1206
• /
• 2006

Plasminogen kringle 5 is a potent inhibitor of endothelial tell proliferation like an endogenous angiogenesis inhibitor, angiostatin consisting of plasminogen kringles 1-4. In this study, we produced the recombinant protein of plasminogen kringle 5 (PK5) employing an Pichia expression system and examined its. effect on~endothelial cell migration and its possible inhibitory mechanism. PK5 was expressed in Pichia pastoris GS115 by fusion of the cDNA spanning from Thr456 to Phe546 to the secretion signal sequence of a-factor prepro-peptide. After methanol induction, the secreted PK5 was purified by using S-spin column. SDS-PACE analysis of the purified protein showed one major band of approximately 10kDa. In in vitro migration assays, the purified protein inhibited dose-dependently the migration of human umbilical endothelial cells (HUVECs) induced by basic fibroblast growth factor (bFGF) or vascular endothelial growth factor (VEGF) with an $IC_{50}$ of approximately 500nM. Accordingly, it inhibited bfGF-stimulated extracellular signal-regulated kinase 1/2 (ERK1/2) phosphorylation in HUVECs at 500nM. In addition, it also potently inhibited bFGF-induced cytoskeletal rearrangement of HUVECs. Thus, these results suggest that Pichia-produced PK5 effectively inhibits endothelial cell migration, in part by suppression of ERK1/2 activation and blocking cytoskeleton rearrangement.

• Korean Journal of Microbiology
• /
• v.45 no.4
• /
• pp.291-299
• /
• 2009

To elucidate HtrA3's functional roles in the HtrA3 mediated cellular processes, it is necessary to investigate its biochemical characteristics. In the present study, we constructed the plasmids encoding putative mature HtrA3 proteins (M1-HtrA3 and M2-HtrA3) based on the putative maturation sites of highly homologous HtrA1 and mouse HtrA3. We used the pGEX bacterial expression system to develop a simple and rapid purification for the recombinant HtrA3 protein. Although yields of the mature HtrA3 proteins were slightly low as 10~50 ${\mu}g$/L, the amounts and purity of M1- and M2-HtrA3 were enough to investigate their proteolytic activities. The putative mature HtrA3 proteins have proteolytic activity which could cleave $\beta$-casein as an exogenous substrate. We compared the proteolytic activity between the HtrA family, HtrA1, HtrA2, and HtrA3. The cleavage activity of HtrA3 and HtrA2 were 2 folds higher than that of HtrA1, respectively. Our study provides a method for generating useful reagents to identify natural substrates of HtrA3 in the further studies.

• Cartoon and Animation Studies
• /
• s.39
• /
• pp.297-320
• /
• 2015

Modern technology in the development and convergence of art brought the rapid development of the media arts actively accommodating 'media art' digital media. It has to go to expand the area and limitations as a basis of communication in the 21st century, a new artistic creativity, through the gathering of the existing chain of modern digital mobile technologies and new genre public art that is based on interactive communication break down the boundaries between sectors It has evolved into. This fusion of high technology and the art of mobile interactive media art is to overcome the limitations of time and physical space, as an active subject of interaction and participation, and expanding the range of experiential communications, such as art and science, cultural industries giving provides a flexible platform for a variety of applications. This study presents an expanded paradigm of the new communication and interactive media that define the reporting year review through the literature on art, experiential mobile communications through a case study of mobile interactive media art that is used as a medium of artistic expression the interactive effect was analyzed as a possible new public art. Convergence and interactivity, mobile interactive media art as a buzzword experience has proposed a new approach to high-tech and art and meet the new sensibility of our life and communication, unlimited possibilities worthy of the contemporary trend of convergence and consilience with a new art genre is expected to continued to evolve and develop.